PPT-Restriction Enzymes Restriction Modification System

Cutting DNA molecules Before 1970s there was no method of cleaving DNA at the specific points All the available methods for fragmenting DNA were nonspecific Mechanical shearing. Recombinant DNA Technology. T. echnique that allows DNA to be . combined. from different sources. DNA code universal. Foundation of genetic engineering. Review. What are enzymes? What function do they serve?. Restriction Enzymes. General Genetics. Objectives:. Introduce the students to digest genomic DNA by restriction . endonucleases. .. Observe the results of digestion on . agarose. gel electrophoresis.. Stephen. . Halford. DNA-Proteins Interactions Unit,. Department of Biochemistry,. Why study the . enzymology. of Type II restriction enzymes?. . Enzyme specificity. . c.f. . aminoacyl. . tRNA. . IMPORTANCE. OF ENZYMES. Enzymes are biological catalysts that:. . I. ncrease. . reaction rates . by lowering the amount of energy needed for the reaction to occur. 100-1000x . faster. !!!. Activation Energy. 1. What are enzymes?. Each enzyme has only one specific reaction it can perform. . 2. How many different reactions can a single enzyme perform?. Yes, the same enzyme molecule can be used over and over. Part 1. . Chapter 3. Learning outcomes. You should be able to:. explain how enzymes work. describe and explain the factors that affect. enzyme activity. use V. max. and K. m. to compare the affinity of different enzymes for their substrates. Enzyme & Substrate. An enzyme is a globular protein which acts as a biological catalyst by speeding up the rate of chemical reactions. . Enzymes are . NOT. changed or consumed by the reactions they catalyze and thus can be reused. . Vocabulary. Karyotype. Autosome. Sex chromosome. Nondisjunction. Monosomy. Trisomy. Pedigree. Carrier. Restriction enzyme. Restriction site. Restriction fragment. Sticky ends. Recombination. Gel electrophoresis. Objectives. -Understand how Restriction Enzymes digest DNA.. -Know how to construct a . pAMP. . (plasmid) or gel.. -Given the size of fragments, gel, know how to construct a restriction map.. -Given a restriction map know how to construct a gel.. What are restriction endonucleases (REs)?. How can REs be used to identify DNA molecules?. How can I find RE recognition sites in the . MET. plasmids? . Restriction endonucleases are part of a bacterium’s defense against invaders. Restriction enzymes. Polymerase enzyme. Strand-displacing polymerases. Helicase enzymes. Discovery and Obtainability:. Most enzymes are proteins discovered in cells.. But . DNAzymes. were . descovered. are. . working. ?. Restriction. Enzymes. Natural . occurrence. . of. . restriction. . enzymes. . is. in . bacteria. . for. . protecting. . them. . against. a . virus. . attack. . by. . Presented . by. Ms. . P. . . H. . Giri. Department of Microbiology. Deogiri . College, Aurangabad. B.Sc. T. Y. . Semester VI. Paper No. XIX. Recombinant DNA Technology. Ms. . Priyanka. H. . Giri. DNA manipulating enzymes:. the isolation of the restriction . endonuclease. from. E. coli K (. Meselson. & Yuan 1968). . It was evident . that the restriction . endonucleases. from . E. coli K . and . E. coli B were important examples of proteins.